The returning Neymar could not inspire Brazil to victory as hosts Colombia ended their perfect start to World Cup qualifying with a 0-0 draw in Barraquilla on Sunday.
The Selecao had won their first nine qualifiers, scoring 22 goals to lead the single South American table by eight points before kick-off.
Coach Tite's side are still almost certain to qualify for next year's finals in Qatar as they maintain a 13-point lead over fifth-placed Colombia, with the top four progressing automatically.
Copa America winners Argentina could cut Brazil's lead to six points if they beat Uruguay later on Sunday.
Brazil started brightly and dominated the opening quarter as Neymar thrashed a fierce drive that forced David Ospina into a diving parry at his near post.
Lucas Paqueta then stung the goalkeeper's palms from distance and moments later should have put the visitors in front.
Neymar's cute reverse pass put Paqueta in one-on-one with Ospina but on the stretch he poked the ball wide.
Colombia were struggling to create much and their best first half moments came from set-pieces.
Juan Quintero curled a free-kick high and wide having earlier crossed for Yerry Mina to head wide.
Neymar was at the heart of much of Brazil's attacks but when he teed up a teammate on the edge of the area, it was defensive midfielder Fred who appeared on the scene, and it was no surprise to see him blaze over the bar from 18 yards.
Colombia grew more adventurous in the second half and Brazil goalkeeper Alisson tipped a fierce Mateus Uribe drive from distance over the bar.
The Liverpool stopper was called into action again soon afterwards to deny Quintero, while Radamel Falcao had the ball in the net 20 minutes from time but it was disallowed for a foul on Militao.
Substitute Raphinha was at the heart of Brazil's best moments after the break.
He forced Ospina into a save with his left-foot curler from the right but then picked out fellow substitute Antony with a perfect in-swinging delivery, only for Ospina to make a brilliant reaction save from point-blank range.
Colombia almost had the last word but Santos Borre blazed well over from the edge of the area.
It was a good day for the bottom two teams in the group as Bolivia beat Peru 1-0 and Venezuela defeated Ecuador 2-1.
Ramiro Vaca's goal eight minutes from time gave Bolivia victory despite having fellow substitute Henry Vaca sent off on 75 minutes.
Goals from Darwin Machis and Eduard Bello either side of a penalty from Enner Valencia were enough to give Venezuela a rare win.